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Spectroscopy

Spectroscopy is the analysis of the interaction between matter and any portion of the
electromagnetic spectrum. Traditionally, spectroscopy involved the visible spectrum of light, but
X-ray, gamma, and UV spectroscopy also are valuable analytical techniques. Spectroscopy can
involve any interaction between light and matter, including absorption, emission, scattering, etc.

Data obtained from spectroscopy is usually presented as a spectrum (plural: spectra) that is a plot
of the factor being measured as a function of either frequency or wavelength. Emission spectra
and absorption spectra are common examples.
Although chemical test and elemental analysis are useful but they are time consuming and
uncertain, sometimes required a large amount of sample and are destructive. today we can obtain
a great deal of information directly from molecules by the spectroscopic method. Compare to the
chemical analysis, spectroscopic methods are faster and more accurate, require less sample, and
are usually non destructive.

We can define spectroscopy as an area of study that probes molecular structure with
electromagnetic radiation. The instruments used in such studies are called spectrometers. Teh
merits of spectroscopic technique over the classical techniques are as follows:

1. Spectroscopic techniques are quick and very sensitive.

2. Spectroscopic technique requires very small quantity of sample for investigation
compare to classical techniques.
3. These techniques are reliable and reproducible result and provide permanent record of
observation in the form of spectra.
4. The material can be retrieved unchanged after the investigation and can be reuse for other
studies, except in case of mass spectroscopy.
5. Continuous monitoring of changes taking place is possible in spectroscopic techniques.

Types of Spectroscopy

1. Flame Spectroscopy
2. Visible spectroscopy
3. Ultraviolet spectroscopy
4. Infrared spectroscopy
5. Raman Spectroscopy
6. Nuclear magnetic resonance spectroscopy
7. Photoemission spectroscopy

The Electromagnetic Spectrum

Physicists classify light waves by their energies (wavelengths). Labeled in increasing energy, we
might draw the entire electromagnetic spectrum as shown in the figure below:
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The Electromagnetic Spectrum. Notice how small the visible region of the spectrum is,
compared to the entire range of wavelengths.

Notice that radio, TV, and microwave signals are all light waves, they simply lie at wavelengths
(energies) that your eye doesn't respond to. On the other end of the scale, beware the high energy
UV, x-ray, and gamma-ray photons! Each one carries a lot of energy compared to their visible-
and radio-wave brethren. They're the reasons you should wear sunblock, for example.

When we look at the Universe in a different "light", i.e. at "non-visible" wavelengths, we

probe different kinds of physical conditions -- and we can see new kinds of objects! For
example, high-energy gamma-ray and X-ray telescopes tend to see the most energetic dynamos
in the cosmos, such as active galaxies, the remnants from massive dying stars, accretion of
matter around black holes, and so forth. Visible light telescopes best probe light produced by
stars. Longer-wavelength telescopes best probe dark, cool, obscured structures in the Universe:
dusty star-forming regions, dark cold molecular clouds, the primordial radiation emitted by the
formation of the Universe shortly after the Big Bang. Only through studying astronomical
objects at many different wavelengths are astronomers able to piece together a coherent,
comprehensive picture of how the Universe works!

General Types of Spectra

Typically one can observe two distinctive classes of spectra: continous and discrete. For a
continuous spectrum, the light is composed of a wide, continuous range of colors (energies).
With discrete spectra, one sees only bright or dark lines at very distinct and sharply-defined
colors (energies). As we'll discover shortly, discrete spectra with bright lines are
called emission spectra, those with dark lines are termed absorption spectra.

Continuous Spectra

Continuous spectra arise from dense gases or solid objects which radiate their heat away through
the production of light. Such objects emit light over a broad range of wavelengths, thus the
apparent spectrum seems smooth and continuous. Stars emit light in a predominantly (but not
completely!) continuous spectrum.

Discrete Spectra

Discrete spectra are the observable result of the physics of atoms. There are two types of discrete
spectra, emission (bright line spectra) and absorption (dark line spectra). Let's try to understand
where these two types of discrete spectra.
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Emission Spectra

When light emitted by a substance is passed through a prism we obtain certain lines, which are
called emmission spectra.

In the diagram below, a hydrogen atom drops from the 2nd energy level to the 1st, giving off a
wave of light with an energy equal to the difference of energy between levels 2 and 1. This
energy corresponds to a specific color, or wavelength of light -- and thus we see a bright line at
that exact wavelength! ...an emission spectrum is born, as shown below:

An excited Hydrogen atom relaxes from level 2 to level 1, yielding a photon. This results in a
bright emission line.

Absorption Line Spectra

When light is passed through substances, certain portion of light is absorbed by it. Missing
wavelength leaves dark lines or bands at their place. The spectrum obtain is called absorption
spectra. A dark-line absorption spectrum is born, as shown below:

A hydrogen atom in the ground state is excited by a photon of exactly the `right' energy needed
to send it to level 2, absorbing the photon in the process. This results in a dark absorption line.

1. Ultra-violet (U.V) Spectroscopy

The Principle of UV-Visible Spectroscopy is based on the absorption of UV light and visible
lights by some chemical compounds, which produces the distinct spectra. The molecules which
have bonding and nonbonding electrons can absorb energy in the form of UV or visible light to
excite these electrons to higher anti-bonding molecular orbitals. The term Spectroscopy is based
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on the interaction between light and matter, the matter absorbs the light and it undergoes
excitation and de-excitation, and results in the production of the spectrum.

Whenever the matter absorbs any type of UV- radiation, the electrons present in it undergo
excitation. Because of this process, the electrons become excited in the state i.e., they jump from
the ground state (the state of energy in which a small amount of energy is associated) to an
excited state (the energy state in which a large amount of energy is associated with it). The
interesting fact about the difference of ground state and excited state of the electron is always
equal to the amount of UV radiation or visible radiation absorbed by it.

UV spectroscopy obeys the Beer-Lambert law, which states that: when a beam of
monochromatic light is passed through a solution of an absorbing substance, the rate of
decrease of intensity of radiation with thickness of the absorbing solution is proportional to
the incident radiation as well as the concentration of the solution.
According to this law, the greater number of absorbing molecules with the ability to absorb light
of a specific wavelength, the greater the extent of absorption of the radiation.
The expression of Beer-Lambert law is-
A = log (I0/I) = ECL
Where, A = absorbance
I0 = intensity of light incident upon sample cell
I = intensity of light leaving sample cell
C = molar concentration of solute
L = length of sample cell (cm)
E = molar absorptivity
From the Beer-Lambert law it is clear that greater the number of molecules capable of absorbing
light of a given wavelength, the greater the extent of light absorption. This is the basic principle
of UV spectroscopy.
The range of electronic spectroscopy is 200-800nm. This range consists of two parts, 200-400nm
is the range of ultraviolet radiations whereas 400-800nm is the range of visible light.

Instrumentation of UV Spectroscopy
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Learn to gain something

Instrumentation and working of the UV spectrometers can be studied simultaneously. Most of

the modern UV spectrometers consist of the following parts-
Light Source- Tungsten filament lamps and Hydrogen-Deuterium lamps are most widely used
and suitable light source as they cover the whole UV region. Tungsten filament lamps are rich in
red radiations; more specifically they emit the radiations of 375 nm, while the intensity of
Hydrogen-Deuterium lamps falls below 375 nm.

Monochromator- Monochromators generally composed of prisms and slits. The most of the
spectrophotometers are double beam spectrophotometers. The radiation emitted from the
primary source is dispersed with the help of rotating prisms. The various wavelengths of the light
source which are separated by the prism are then selected by the slits such the rotation of the
prism results in a series of continuously increasing wavelength to pass through the slits for
recording purpose. The beam selected by the slit is monochromatic and further divided into two
beams with the help of another prism.

Sample and reference cells- One of the two divided beams is passed through the sample
solution and second beam is passé through the reference solution. Both sample and reference
solution are contained in the cells. These cells are made of either silica or quartz. Glass can't be
used for the cells as it also absorbs light in the UV region.
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Detector- Generally two photocells serve the purpose of detector in UV spectroscopy. One of
the photocell receives the beam from sample cell and second detector receives the beam from the
reference. The intensity of the radiation from the reference cell is stronger than the beam of
sample cell. This results in the generation of pulsating or alternating currents in the photocells.

Amplifier- The alternating current generated in the photocells is transferred to the amplifier. The
amplifier is coupled to a small servometer. Generally current generated in the photocells is of
very low intensity, the main purpose of amplifier is to amplify the signals many times so we can
get clear and recordable signals.

Recording devices- Most of the time amplifier is coupled to a pen recorder which is connected
to the computer. Computer stores all the data generated and produces the spectrum of the desired
compound.
Application:

1. Detection of functional groups- UV spectroscopy is used to detect the presence or absence of

chromophore in the compound. This is technique is not useful for the detection of chromophore
in complex compounds. The absence of a band at a particular band can be seen as an evidence
for the absence of a particular group. If the spectrum of a compound comes out to be transparent
above 200 nm than it confirms the absence of –
a) Conjugation b) A carbonyl group c) Benzene or aromatic compound d) Bromo or iodo atoms.

2. Detection of extent of conjugation- The extent of conjugation in the polyenes can be detected
with the help of UV spectroscopy. With the increase in double bonds the absorption shifts
towards the longer wavelength. If the double bond is increased by 8 in the polyenes then that
polyene appears visible to the human eye as the absorption comes in the visible region.

3. Identification of an unknown compound- An unknown compound can be identified with the

help of UV spectroscopy. The spectrum of unknown compound is compared with the spectrum
of a reference compound and if both the spectrums coincide then it confirms the identification of
the unknown substance.

4. Determination of configurations of geometrical isomers- It is observed that cis-alkenes absorb

at different wavelength than the trans-alkenes. The two isomers can be distinguished with each
other when one of the isomers has non-coplanar structure due to steric hindrances. The cis-
isomer suffers distortion and absorbs at lower wavelength as compared to trans-isomer.

5. Determination of the purity of a substance- Purity of a substance can also be determined with
the help of UV spectroscopy. The absorption of the sample solution is compared with the
absorption of the reference solution. The intensity of the absorption can be used for the relative
calculation of the purity of the sample substance.
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Some Terms:
Chromophore- Chromophore is defined as any isolated covalently bonded group that shows a
characteristic absorption in the ultraviolet or visible region (200-800 nm). Chromophores can
be divided into two groups-
a) Chromophores which contain p electrons and which undergo π to π* transitions. Ethylenes and
acetylenes are the example of such chromophores.
b) Chromophores which contain both p and nonbonding electrons. They undergo two types of
transitions; π to π* and nonbonding to π*. Carbonyl, nitriles, azo compounds, nitro compounds
etc. are the example of such chromophores.

Auxochromes- An auxochrome can be defined as any group which does not itself act as a
chromophore but whose presence brings about a shift of the absorption band towards the longer
wavelength of the spectrum. –OH,-OR,-NH2,-NHR, -SH etc. are the examples of auxochromic
groups.

Infrared Spectroscopy

Basic Principle
The atoms in a molecule are in a state of constant vibratory and rotatory motion. The molecular
vibrations are the two types (i) stretching and (ii) bending.
A strectching vibration is a rhythmical movement along the bond axis such that interatomic
distance changes. In bending vibration, the bond angle change. Only these vibrations that result
in a rhythmical change in the dipole moment are observed in infrared. Absorption of infrared
light can occur only if the dipole moment of the molecule differs in two vibrational levels.
Thus, there are a number of vibration and rotational changes in the molecule. On absorption
light, the molecules are excited from lower vibration and rotational levels to higher ones. For
affecting such transitions, frequencies in the infrared range are good enough. Molecules will
absorb such frequencies as are needed to excite molecules from lower vibration levels to
permitted higher levels. Absorption of energy by a bond increases the amplitude of vibrations.
Every bond and every functional group has a specific absorption frequency which is calculated to
excite the molecule to higher vibrational or rotational level.
A graph between the amount of absorption (or emission) of infrared light against the frequency
(or wavelength) of this light is called infrared (IR) spectrum.
The two important areas of preliminary examination of a spectrum are the region 400
cm' and 909-650cm". The high frequency region is called functional region. The lack of strong
absorption band in 909-650cmgenerally indicates a non aromatic structure. The intermediate
portion of the spectrum 1300-909cm" is usual called finger print region. The absorption pattern
here is very complex. It is because many bending and stretching vibrations. As there are many
bending vibration in molecule then stretching vibration this region of the spectrum is rich in
absorption bands and shoulders. This portions of the spectrum is used to compare the spectra of
various compounds.

IR absorption bands of some important functional group

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Functional group Type of Compound Frequency

>C=0 aldehydo, ketone 1690 - 1760
carbohylic acids, estes
- ОН Monomeric alcohols and 3610 - 36400
phenol
- ОН H-bonded alcohol and 3200 - 3600 (broad)
phenol
- ОН H bonded carboxylic 2500 - 3000 (broad)
acid
- NH amines 3300 - 3500
-NO2 nitro compound 1515 - 1560
-C=N nitriles 2210 - 2260

Application of IR Spectroscopy:
(i) Identification of the functional group:
Every functional group absorbs IR radiation in a particular range irrespective of the compound in
which such group is present. For example, a carbonyl group gives IR absorption band in the
range of 1650 - 1750cm-1. Hence by observing the absorption frequency, we can tell something
about the functional group in the compound.
(ii) Determination of structure:
Investigation of the IR spectra gives us information regarding functional groups present in the
molecule. Further, the IR spectra in the fingerprint region gives us an insight into various other
groups and their relative positions. The two studies together help us to arrive at the exact
compound.
(iii) Testing purity of a compound:
Generally, a pure compound has well defined and sharp absorption frequencies whereas the
spectra of impure compound has diffused lines due to additional impurity materials present in the
sample. IR spectra of many pure compounds are available in literature. We can compare the IR
spectra of the unknown sample with that of the pure sample. If the two tally exactly, the sample
is pure, otherwise not.
(iv) Following progress of a reaction:
During the course of reaction, many intermediate compounds are formed which change into the
products. These intermediate compounds can be identified by IR spectroscopy. IR
spectrophotometer records the spectra and by interpreting them, we can follow the reaction.
Thus, the mechanism of the reaction can be proposed.
[Link].1. What type of structural information is obtained from IR spectroscopy? Explain giving
example.
Ans: (i) Identification of the functional group:
Every functional group absorbs IR radiation in a particular range irrespective of the compound in
which such group is present. For example, a carbonyl group gives IR absorption band in the
range of 1650 - 1750cm-1. Hence by observing the absorption frequency, we can tell something
about the functional group in the compound.
(ii) Determination of structure:
Investigation of the IR spectra gives us information regarding functional groups present in the
molecule. Further, the IR spectra in the fingerprint region gives us an insight into various other
groups and their relative positions. The two studies together help us to arrive at the exact
compound.
(iii) Testing purity of a compound:
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Generally, a pure compound has well defined and sharp absorption frequencies whereas the
spectra of impure compound has diffused lines due to additional impurity materials present in the
sample. IR spectra of many pure compounds are available in literature. We can compare the IR
spectra of the unknown sample with that of the pure sample. If the two tally exactly, the sample
is pure, otherwise not.
(iv) Following progress of a reaction:
During the course of reaction, many intermediate compounds are formed which change into the
products. These intermediate compounds can be identified by IR spectroscopy. IR
spectrophotometer records the spectra and by interpreting them, we can follow the reaction.
Thus, the mechanism of the reaction can be proposed.
Example: An aromatic compound having molecular formula C8H8O shows IR absorption band
at 1690cm-1 but no band at 3300cm-1. From this data we can predict the structure of an
compound.
An absorption band at 1690cm-1 shows that a carbonyl (>C=O) group is present in the molecule.
Absence of band at 3300cm-1 shows that molecule does not contain an alcoholic group (-OH)
group. Hence the structure of the compound is:

C CH3

Mass Spectroscopy
Mass spectroscopy involves the bombardment of molecules of substance by energetic electrons
and analyzes the charged particle and fragments. The instruments used for this purpose is called
mass spectrometer. Mass spectroscopy is one of the widely used spectroscopic techniques used
for the structure elucidation of organic compound.
Basic Principle:
In mass spectroscopy, the molecule of a substance is bombarded with medium energetic
electrons. The energetic electrons knock out generally one most loosely bound electron in the
molecule. In rare cases, two electrons may be removed. This process produces cations or
molecular ions. The molecular ion being energetic is fragmented. Further in steps to produce
smaller ions called daughter ions or fragmented ions. Each of the daughter ions has certain m/e
ratio i.e, ratio of mass to change of the ions.
The molecular ion and daughter ions having different m/e ratio are separated by deflection in a
variable magnetic field according to their mass and charge and generate a current (ion current) at
the collector in proportion to their relative abundance.
A mass spectrometer performs three essential functions:
1. Molecules are bombarded with a stream of high energy electrons converting some of the
molecules in to ions. These ions are accelerated by electric field.
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2. The accelerated ions are separated according to their m/e ratio in a magnetic or electric
field.
3. Ions with a particular m/e ratio are detected by a device which is to count the number of
ions which strike it. The detector output is amplified and fed into the recorder. A signal is
obtained corresponding to each m/e ratio value. The intensity of signal is proportional to
the number of ions giving rise to the signal. A graph between intensity relative abundance
and m/e values of the ion is called mass spectrum.
The most intense peak is referred to as base peak and its intensity is taken as 100. The intensity
of other ions are measured with respect to base peak.

e- -2e-
There are two types of fragments: M+

Application of mass spectroscopy:

1. Identification of substance:
Mass spectrum is highly characteristic of a substance. The mass spectra of two identical samples
superimpose exactly on each other. By comparing the spectra of an unknown substance with
those of known samples, it is possible to identify the unknown samples.
2. Determination of molecular mass and formula:
Molecular mass of a compound is given m/e value of the molecular ion in the mass spectrum.
M + e- → M+ + 2e-
All that has to be done in interpreting the mass spectrum is to identify the M* peak (In most
cases, it is the base peak). The molecular mass obtained in this way is the mass corresponding to
common isotopes of the elements.
Once the molecular mass has been obtained, it is not difficult to arrive at molecular formula.
3. Determination of molecular structure:
Mass spectrum gives us various value of m/e which corresponds to various fragments of the
molecular ion. By establishing, the nature of different fragments, we can know about the
structure of the compound by joining those fragments together.
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Nuclear Magnetic Resonance (NMR) Spectroscopy

The magnetic properties of nuclei in most of the organic compound are the basis of nuclear
magnetic resonance spectroscopy.

If a nucleus like proton is placed in an external magnetic field, the magnetic moment of proton
will be oriented either with or against the external field. Out of two orientations the one with the
applied field is more stable (smaller energy) then one against the applied field. The difference in
energies of the two orientations is designated by ∆E. Its magnitude varies with the magnitude of
applied field. The difference is however small and lies within the range of radio frequency region
of electromagnetic spectrum. Thus, if we desire to flip or shift the nucleus from lower energy
state to higher energy state, then amount of ∆E will have to be absorbed by the nucleus.

In organic chemistry, we are most interested to the proton as the nuclei, as hydrogen is
constituent of almost every organic compound. The particular branch of NMR spectroscopy,
where the nucleus is a proton is called proton magnetic resonance (PMR). However, principally,
NMR and PMR spectroscopy are same.

A PMR spectrum can be recorded by placing the substance containing hydrogen nuclei in
magnetic field of constant strength and passing radiation of changing frequency through the
substance and noting the frequency at which the absorption of energy corresponding to flipping
of proton from lower to higher energy state takes place. However for ease(easy) of operation, the
frequency of radiation is kept constant(at 40,60 or 100 MHz) and the strength of magnetic field
is changed.
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On plotting, a graph of absorption of energy versus magnetic field strength at constant frequency
of radiation, a signal is obtained which signified the absorption of energy at particular field
strength.

absorption
of radiation

The following information regarding structure of molecule can be obtained from NMR
spectrophotometer:

1. The position of the signal:

It tells us something about the electronic environment of each kind of proton.

2. The number of signals:

This tells us how many different kind of proton are present in the molecule.

3. The splitting of a signal in to several peaks

This tells us about the environment of a protons of with respect to other, nearby protons.

Application of NMR spectroscopy:

1. Iidentification of functional group:

Every functional group gives a characteristic signal in the NMR spectrum. By studying the
chemical shift of the compound, it becomes possible to establish what kind of the functional
group is present in the given compound.

2. Structure of an unknown compound:

It is possible to elucidate the structure of an unknown compound from the NMR studies. This is
because protons under different environment give different chemical shifts. By studying
doublets, triplets, and multiples, it is possible to place hydrogen at appropriate place in the
formula and hence to establish the structure.

3. Comparison of two compounds:

NMR spectra is like fingerprint of a compound. Two compound showing same NMR spectra
must be structurally identical.
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Study well

Happy Bijaya Dashami & Suvha Dipawoli